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Kandelia Candel</Emphasis> View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Xiamen University Institutional Repository Hydrobiologia 479: 75–81, 2002. 75 © 2002 Kluwer Academic Publishers. Printed in the Netherlands. Growth and physiological responses of Kandelia candel and Bruguiera gymnorrhiza to livestock wastewater Y. Ye 1,2 &NoraF.Y.Tam2 1Marine Biotechnology Key Lab, Ningbo University, Ningbo, Zhejiang, China E-mail: [email protected] 2Department of Biology and Chemistry, City University of Hong Kong, Hong Kong Received 15 March 2001; in revised form 6 March 2002; accepted 31 March 2002 Key words: Kandelia candel, Bruguiera gymnorrhiza, mangrove, livestock wastewater, salinity Abstract Growth and physiological responses of two mangrove species (Kandelia candel and Bruguiera gymnorrhiza)to livestock wastewater under two salinity conditions (seawater with salinity of 30þ and freshwater) were examined in greenhouse pot-cultivation systems for 144 days. Wastewater treatment significantly enhanced growth of Kandelia candel and Bruguiera gymnorrhiza in terms of stem height, stem basal diameter, leaf production, maximum unit leaf area and relative growth rate. Wastewater discharges and salinity levels did not significantly change biomass partitioning of Kandelia candel, however, more biomass of Bruguiera gymnorrhiza was allocated to leaf due to wastewater discharges. In Bruguiera gymnorrhiza, contents of chlorophyll a and chlorophyll b increased with wastewater discharges but such increase was not observed in Kandelia candel. On the other hand, livestock wastewater increased leaf electric conductance in Kandelia candel but not in Bruguiera gymnorrhiza. The per- oxidase activity in stem and root of Kandelia candel under both salinity conditions increased due to wastewater discharges, while the activity in root of the treated Bruguiera gymnorrhiza seedlings decreased under freshwater condition but increased at seawater salinity. The superoxide dismutase activity in treated Bruguiera gymnorrhiza decreased but did not show any significant change in Kandelia candel receiving livestock wastewater. Introduction removal should be selected. The species should also be tolerant to wastewater stress. So growth and physiolo- Natural and constructed wetlands including man- gical responses of the wastewater treated plants should groves have been considered to be low-cost and effect- be an important consideration when selecting the ap- ive wastewater treatment systems and are especially propriate species for wastewater treatment. efficient in nutrient removal (Clough et al., 1983; Cor- Mangroves are naturally distributed in areas with a bitt & Bowen, 1994; Breaux et al., 1995). Nutrients, wide range of water salinity, from nearly freshwater especially nitrogen and phosphorus are often deficient to seawater (Lin, 1999). Habitat salinity may influ- in coastal inter-tidal areas such as mangrove wetlands ence the N uptake by mangrove plants, and affect (Boto & Wellington, 1983; Clough et al., 1983; Lin, their growth and physiological responses to wastewa- 1999). Therefore, mangrove wetlands are particularly ter addition. Naidoo (1990) suggested that growth suitable for the treatment of wastewater rich in nu- enhancement of Bruguiera gymnorrhiza in response to trients. In a mangrove ecosystem, mangrove plant is N addition occurs only at a low salinity. The present the most important component. If mangroves were investigation therefore aims to compare growth and to be used as the treatment system for nutrient-rich physiological responses of two dominant mangrove wastewater such as livestock sewage, suitable man- plant species in Hong Kong SAR and South-east Asia, grove plant species that is most efficient in nutrient Bruguiera gymnorrhiza and Kandelia candel,tolive- 76 Table 1. Characteristics of livestock wastewater, tap water and seawater used in this experiment Parameter Wastewater Tap water Seawater pH 7.24 (7.00∼7.35) 7.64 (7.10∼8.03) 8.01 (7.32∼8.26) − Conductivity (ms cm 1) 1.39 (1.14∼1.77) 0.22 (0.18∼0.23) 36.60 (34.83∼38.49) − TOC (mg l 1) 65.9 (48.1∼88.3) 2.3 (0.5∼5.4) 5.2 (2.1∼7.0) + −1 NH4 -N (mg l ) 36.1 (21.2∼58.7) 0.3 (0.0∼1.0) 1.9(0.0∼5.9) − −1 NOx -N (mg l ) <0.05 <0.05 <0.05 3− −1 PO4 -P (mg l ) 53.7 (22.5∼94.6) 0.2 (0.0∼0.5) 0.2(0.0∼0.7) Mean and range (in brackets) values of 9 samples are shown. stock wastewater. The effect of salinity on these re- 0.64 ± 0.05 cm and the stem height was 25.2 ± 3.9 sponses was also evaluated. This work is of significant cm (n=36). importance to the selection of the most suitable plant During the treatment period, each pot was daily species of mangrove wetlands to treat wastewater rich watered with 300 ml treatment liquid. For each plant in nutrients. species, the following four treatments, each in triplic- ate and lasted for 144 days, were set up: (1) F, each pot was irrigated with tap water every day, (2) FW, each pot was irrigated with wastewater in Materials and methods every three days and tap water in every other day, (3) S, each pot was irrigated with artificial seawater Experimental design (salinity of 30þ, prepared by dissolving a commer- cial salt purchased from Instant Ocean, Aquarium In April 1997, mature propagules of Bruguiera gym- Systems, Inc., Mentor, Ohio) every day, norrhiza were collected from Mai Po Mangrove ◦ ◦ (4) SW, each pot was irrigated with wastewater in Nature Reserve (114 05 E, 22 32 N) in Hong every three days and seawater in every other day. Kong SAR and planted into plastic pots (18 cm in diameter and 20 cm in height), with four individuals Growth and biomass partitioning analysis per pot, in a greenhouse. In November 1998, 1.5 years old Kandelia candel seedlings were transplanted from At the beginning and at the end of the experiment, the Wong Chuk Wan mangrove swamp of Hong Kong into stem basal diameter (D) at the first stem node and the greenhouse pots with three individuals per pot. Each stem height (H) (excluding hypocotyl) of each plant pot contained 4 kg soils freshly collected from Sai were measured. At the end of the experiment, biomass Keng mangrove swamp, HKSAR. The soil is a loamy- partitioning was determined by 105 ◦C dry weights sandy soil with 73.11%, 15.93% and 10.96% of sand, of shoot (leaf and stem) and root of each individual silt and clay particles, respectively. Each pot was daily plant. Hypocotyl was excluded in either shoot or root irrigated with 300 ml tap water and excess water was biomass measurement, mainly because the weight of able to drain freely from the six draining holes (0.6 cm hypocotyl did not change during the experiment. Data in diameter) at the bottom by gravitational force. on total biomass per pot (B), average stem basal dia- On 3 March 1999, 12 pots of B. gymnorrhiza and meter per pot (D) and average stem height per pot 12 pots of K. candel were used for treating livestock (H) collected at the end of the experiment were used wastewater. The livestock sewage was collected from to fit the non-destructive allometric equations sugges- Ta Kwu Ling Pig Farm of Hong Kong SAR after be- ted by Snedaker & Snedaker (1984). The calculated ing primarily treated by sedimentation and secondarily equations for the two species were as follows: treated by aeration. The wastewater had high P content + 3− B. gymnorrhiza : log B = 0.9998 + and low NH4 -N/PO4 –P ratio of 1:1.5 (Table 1). The stem basal diameter and stem height of Bruguiera 0.5416 log (D2H)(n = 12, p < 0.01) gymnorrhiza at the start of the experiment were 0.82 = + ± 0.08 cm and 24.2 ± 3.2 cm, respectively (n=48), K. candel:logB 0.6108 0.6450 while the basal stem diameter of Kandelia candel was log (D2H)(n = 12,p <0.01) 77 The initial biomass per pot was then estimated accord- ing to the equations. The relative growth rate (RGR) was calculated as ln B –lnB RGR = 2 1 . t2 − t1 B1 and B2 were total biomass at the beginning (t1)and at the end (t2) of the experimental period (Hunt, 1978). Physiological analysis About 0.1 g fresh tissues of the third pair of leaves from the top were ground in cold mortar with 10 ml 80% acetone. The homogenate was centrifuged at 10 000× g for 3 min. The contents of chlorophyll a, chlorophyll b, total chlorophyll and total carotenoid were determined according to Lichtenthaler & Well- burn’s method (1983). The methods used to determine leaf electric conductance and root activity were the same as described by Zhang (1990). The methods for extraction and determination of peroxidase (POX) and superoxide dismutase (SOD) activity were similar to those described by Liu & Zhang (1994) with minor modifications. Portions of fresh plant materials were ground and homogenized with five times of 62.5 mmol l−1 phosphate buffer (pH 7.8, including 0.4% polyvinyl pyrrolidone) in an ice bath. The homogenate was centrifuged at 1500 × g for 20 min. The supernatant, the extract of POX and SOD, was stored at 4 ◦C before the enzyme activity was determined. Figure 1. Growth of Kandelia candel (Kc) and Bruguiera gymnor- For the POX activity assay, 20 µl of the enzyme rhiza (Bg) at the end of 144 days treatment (for each species, means extract was diluted to 1 ml with deionized water, with different letters are significantly different at p<0.05 level). mixed with 3 ml combined reagent (consists of 500 −1 ml 0.1 mol l phosphate buffer at pH 7.0, 280 µl was measured. An enzyme unit was calculated as 50% guaiacol and 190 µl hydrogen peroxide).
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